基于UPLC-QqQ-MS/MS同步检测热加工食品中典型晚期糖基化终末产物

doi:10.7506/spkx1002-6630-20200527-332

食品科学 ›› 2021, Vol. 42 ›› Issue (10): 316-321.doi: 10.7506/spkx1002-6630-20200527-332

基于UPLC-QqQ-MS/MS同步检测热加工食品中典型晚期糖基化终末产物

程威威，王霞，张忠飞，刘国琴，陈峰，郑家荣

（1.深圳大学高等研究院，深圳食品产业创新发展研究院，深圳市海洋微生物组工程重点实验室，广东深圳 518060；2.华南理工大学食品科学与工程学院，广东广州 510640）

出版日期:2021-05-25 发布日期:2021-06-02
基金资助:
广东省重点研究计划项目（2019B020212001）；国家自然科学基金青年科学基金项目（32001814）； “十三五”国家重点研发计划重点专项（2016YFD0400204）

Simultaneous Determination of Typical Advanced Glycation End Products in Thermally Processed Foods by Ultra-high Performance Liquid Chromatography-Quadrupole-Tandem Mass Spectrometry

CHENG Weiwei, WANG Xia, ZHANG Zhongfei, LIU Guoqin, CHEN Feng, CHENG Kawing

(1. Shenzhen Key Laboratory of Marine Microbiome Engineering, Institute for Innovative Development of Food Industry, Institute for Advanced Study, Shenzhen University, Shenzhen 518060, China;2. School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, China)

Online:2021-05-25 Published:2021-06-02

摘要/Abstract

摘要： 建立超高效液相色谱-三重四极杆串联质谱法测定热加工食品中羧甲基赖氨酸（Nε-carboxymethyllysine，CML）和羧乙基赖氨酸（Nε-carboxyethyllysine，CEL）含量的同步检测方法。首先脱脂样品经硼氢化钠还原8 h，沉淀蛋白，酸水解后，经Oasis MCX固相萃取小柱净化和富集，以乙腈和含0.1%甲酸的1 mmol/L乙酸铵溶液为流动相梯度洗脱，采用ACQUITY UPLC HSS T3-C18色谱柱分离，多反应监测模式进行定性定量分析。CML和CEL在0.25～500 ng/mL范围内线性良好，相关系数高于0.999，方法检出限分别为8 ng/g和10 ng/g，定量限分别为36 ng/g和40 ng/g。平均回收率分别为96%～103%和94%～107%，相对标准偏差分别为1.48%～2.43%和1.23%～1.84%。利用该方法检测13 种市售热加工食品发现，婴儿肉松和婴儿饼干中CML和CEL含量显著高于烘焙食品和油炸食品（P＜0.05）。结果表明该方法快速、准确、高效，适用于热加工食品中CML和CEL的同步快速检测。

关键词: 超高效液相色谱-三重四极杆串联质谱；热加工食品；羧甲基赖氨酸；羧乙基赖氨酸

Abstract: A rapid and effective method was developed for the simultaneous determination of Nε-carboxymethyllysine (CML) and Nε-carboxyethyllysine (CEL) in thermally processed foods by ultra-high performance liquid chromatography-triple quadrupole-tandem mass spectrometry (UPLC-QqQ-MS/MS). After the defatted samples were reduced by sodium borohydride for 8 h, the protein was precipitated with a mixed solvent of chloroform and methanol (2:1, V/V) and then hydrolyzed with hydrochloric acid. Subsequently, CML and CEL were concentrated and purified on an Oasis MCX SPE column, and separated on ACQUITY UPLC HSS T3-C18 analytical column with gradient elution using a mobile phase consisting of 1 mmol/L aqueous ammonium acetate containing 0.1% formic acid and acetonitrile. The qualitative and quantitative analysis was performed in the positive ionization mode with multiple reaction monitoring (MRM). The results showed that the developed method had a good linear relationship in the concentration of 0.25–500 ng/mL with a correlation coefficient of over 0.999. The limit of detection (LOD) was 8 and 10 ng/g , and the limit of quantitation (LOQ) was 36 and 40 ng/g for CML and CEL, respectively. The average recoveries of CML and CEL at three spiked concentration levels were 96%–103% and 94%–107%, respectively, and the precision, expressed by relative standard deviations (RSD), was 1.48%–2.43% and 1.23%–1.84%, respectively. CML and CEL contents in 13 commercially available thermally processed foods were detected by this method. It was found that the contents of CML and CEL in infant meat floss and biscuits were significantly higher than those in baked and fried foods (P < 0.05). The results indicate that the method established in this study is rapid, precise, and effective for the simultaneous determination of CML and CEL in thermally processed foods.

Key words: ultra-high performance liquid chromatography-quadrupole-tandem mass spectrometry; thermally processed foods; Nε-carboxymethyllysine; Nε-carboxyethyllysine

中图分类号:

TS201.2

程威威，王霞，张忠飞，刘国琴，陈峰，郑家荣. 基于UPLC-QqQ-MS/MS同步检测热加工食品中典型晚期糖基化终末产物[J]. 食品科学, 2021, 42(10): 316-321.

CHENG Weiwei, WANG Xia, ZHANG Zhongfei, LIU Guoqin, CHEN Feng, CHENG Kawing. Simultaneous Determination of Typical Advanced Glycation End Products in Thermally Processed Foods by Ultra-high Performance Liquid Chromatography-Quadrupole-Tandem Mass Spectrometry[J]. FOOD SCIENCE, 2021, 42(10): 316-321.

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基于UPLC-QqQ-MS/MS同步检测热加工食品中典型晚期糖基化终末产物

Simultaneous Determination of Typical Advanced Glycation End Products in Thermally Processed Foods by Ultra-high Performance Liquid Chromatography-Quadrupole-Tandem Mass Spectrometry

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